Respiratory infections have a significant health and economic impact worldwide. Current test products for the diagnosis of respiratory viral infections such as influenza are inadequate for the timely diagnosis needed for successful implementation of antiviral treatment. Unfortunately a test product that can accurately diagnose common respiratory viruses with high confidence in less than 30 minutes directly at point of need has not yet been developed. We propose to develop an innovative device and reagents for the molecular diagnosis of multiple RNA pathogens that will set a new standard for rapid patient care. This application entitled Point of Care Detection of Respiratory Viruses seeks to develop a turnkey solution for the processing, detection, and digital readout of influenza viruses A and B, as well as respiratory syncytial virus (RSV) in 30 minutes. Training requirements for personnel operating the device are minimal. The combination of novel molecular technologies, simple protocol, and affordable multiplex testing capabilities residing in a single, inexpensive device wil enable near point of care diagnostics for a number of serious infectious agents. The research design for achieving this goal includes four processing elements: 1) A one step sample prep that can be completed in a minute, 2) Isothermal amplification of viral RNA in 20 minutes, 3) multiplex sequence specific detection of influenza A, influenza B and RSV including built in standards in a few minutes, and 4) a digital readout of results in electronic form. These steps all occur within a small 4 x 6 inch instrument containing a disposable reagent cartridge. A key factor enabling this game changing technology is the development of a novel enzyme that converts RNA to DNA and isothermally amplifies it in minutes. These attributes have allowed the design of a device that does not use any microfluidics, pumps or valves. The greatly simplified construction significantly reduces the cost. This new technology is ideal for to low resource and battlefield settings, and has long term potential as an over the counter device. The test device and reagents developed as part of this application will be validated in a clinical setting with outside experts. During Phase III of the project we will apply for FDA clearance of the instrument and reagents and commercialize the diagnostic device in the US and other markets around the globe.